Detection of Drug-Induced DNA Hypermethylation in Human Tumor Cells Exposed to Cancer Chemotherapy Agents

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Abstract
A method is described for the quantitative analysis of drug-induced DNA hypermethylation in human tumor cells exposed in vivo or in vitro to DNA synthesis-inhibiting levels of cancer chemotherapy agents. The method relies upon isocratic separation of formic acid hydrolysates of DNA using an Aminex A-9 column converted to the cation form. When combined with pre-incorporation of uridine or deoxycytidine labeled specifically in the 6-position, the method is sensitive in the picomolar range for 5-methyl-cytosine, and is specific for the detection of cytosines methylated during the period of drug-induced DNA synthesis inhibition. The method can be used to quantitate DNA hypermethylation occurring in patient bone marrow or peripheral blood specimens in vivo during chemotherapy.

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